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RLS
GENICON RLS MICROARRAYS
We now provide microarray services to support the
Genicon/Invitrogen ultra-sensitive Resonance Light Scattering microarray
platform for gold and silver particle labeling of as little as 1mg total RNA.
Our protocol utilizes amino-allyl
labeling of first strand cDNA followed by conjugation of
monofunctional biotin and fluorescein and then conjugation with avidin-Ag
and anti-fluorescein-Au particles. After archiving, the hybridized
arrays are stable and can stored and rescanned many times.
The HiLight reader
is a white light
scanner that illuminates the array and captures the light scattered by
the gold and silver particles using a CCD camera. Data is stored as a .tiff image. We use
ArrayVision software to grid and analyze the data.
Figure 1.
The Resource has applied an
amino-allyl approach for RLS labeling instead of directly incorporating
the more expensive biotin or fluorescein d-UTP during reverse
transcription. 5ug total HeLa and HL60 RNAs
were amino-allyl labeling during cDNA creation using standard
DeRisi protocols to provide less bias between samples than by
direct incorporation. DNAs were linked to mono-functional biotin or fluorescein and reacted with anti-biotin or anti-fluorescein
tagged gold or silver particles and hybridized against Human 4.6K cDNA array.
Please contact
Irina Tikhonova for
further details about full service projects or slide availability for
RLS approach.
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